Screening method of indigenous group induction quenched bacteria and application thereof
A technology of quorum sensing and screening method, applied in bacteria and other directions, can solve the problems of affecting membrane separation performance, replacement frequency service life, increase MBR operating cost, and high membrane price, so as to reduce membrane fouling, simple and fast production, and prolong replacement cycle. Effect
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Embodiment 1
[0028] A screening method for quorum sensing quenching bacteria according to the present invention, such as Figure 4 As shown, the specific steps are as follows:
[0029] (1) From the activated sludge obtained from the reaction tank of the MBR device in the laboratory test, 1ml of the activated sludge was inoculated on the basic medium with a concentration of 2mM C6-AHL as the only carbon source, and after 5 days of enrichment culture Finally, take 1% of the sample by volume and transfer it to a new basic medium containing C6-AHL for cultivation, and repeat the above steps for 3 times;
[0030] (2) Pick an appropriate amount of cultured bacteria liquid and streak culture on LB solid medium, culture at 30°C for 10-14 hours, repeat the above steps 2-3 times to purify the bacteria, and pick a single colony for storage .
[0031] (3) The basic medium composition in the above steps is:
[0032] Major elements (g / L): NaCl 1.0, KCl 0.5, MgCl 2 ·6H 2 O 0.4, CaCl 2 2H 2 O 0.1, ...
Embodiment 2
[0038] The quorum induction quenching bacteria screened by the present invention is used to degrade the N-acyl homoserine lactone in the activated sludge, comprising the following steps:
[0039] Pick the quorum sensing quenched bacterial colony, inoculate it into LB liquid medium for overnight culture, and cultivate it at a constant temperature of 30°C for 10-14 hours, take the mixed solution and centrifuge it, wash the centrifuged bacteria repeatedly with normal saline, and then use ultrapure water to Bacteria are diluted to a bacterial solution with OD600=1.0;
[0040]Mix the bacterial solution with C6-AHL and C8-HSL at a ratio of 100:1, so that the final concentration of C6-AHL and C8-HSL in the mixed solution is 100ng / mL, and the mixed solution is placed in a constant temperature shaking incubator at 30°C Shake with the condition of 120rpm, and take samples at 10min, 20min, 60min and 120min time points respectively, measure the remaining C6-AHL and C8-HSL concentration in...
Embodiment 3
[0045] Pick colonies of the quorum quenching bacteria and inoculate them into LB liquid medium, culture them in a constant temperature incubator at a constant temperature of 30°C for 10-14 hours, take the mixed solution and centrifuge, wash the centrifuged bacteria repeatedly with physiological saline for 3 times, Discard the supernatant, weigh 0.1g of bacteria (wet weight), embed the bacteria with a PVDF flat film, seal the flat film with epoxy resin and place it in the air to solidify so that it becomes a dumpling-shaped bacterial bag, and the bacterial bag is formed later as image 3 shown.
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